Camptothecin complexes

ABSTRACT

Disclosed are compositions that include a camptothecin and an amorphous cyclodextrin. The camptothecin may be substituted or unsubstituted. Also disclosed are methods of treating undesirable or uncontrolled cell proliferation by administering the inventive compositions. Finally, implants including an implant structure and the inventive composition are disclosed.

CROSS-REFERENCE CROSS REFERENCE TO RELATED APPLICATION

This application is a continuation of U.S. application Ser. No.10/319,783, filed Dec. 13, 2002, which is a continuation of U.S.application Ser. No. 09/539,982, filed Mar. 31, 2000. These applicationsare incorporated herein by reference in its entirety and to whichapplication we claim priority under 35 USC § 120.

BACKGROUND OF THE INVENTION

1. Field of the Invention

The present invention relates to improved formulations for theadministration of certain camptothecin complexes.

2. Description of Related Art

20(S)-camptothecin (CPT), a plant alkaloid, was found to have anticanceractivity in the late 1950's. Wall, M. et al., Plant antitumor agents. I.The isolation and structure of camptothecin, a novel alkaloidal leukemiaand tumor inhibitor from Camptotheca acuminata, J. Am. Chem. Soc. 88:3888-3890, (1966); Monroe E. Wall et al., Camptothecin: Discovery toClinic, 803 Annals of the New York Academy of Sciences 1 (1996). Thesedocuments, and all documents (articles, patents, etc.) cited to herein,are incorporated by reference into the specification as if reproducedfully below. The chemical formula of camptothecin was determined to beC₂₀H₁₆N₂O₄.

CPT itself is insoluble in water. However, during the sixties andseventies the sodium salt of CPT was derived from CPT through opening ofthe lactone ring using a mild base. Clinical trials were then conductedusing this hydrosoluble, sodium salt derivative of CPT (CPT Na+), whichwas administered intravenously. The studies were later abandoned becauseof the high toxicity and low potency of CPT Na+. Gottlieb, J. A., etal., Preliminary pharmacological and clinical evaluation of camptothecinsodium salt (NSC 100880), Cancer Chemother. Rep. 54:461-470 (1979);Muggia, F. M., et al., Phase I clinical trials of weekly and dailytreatment with camptothecin (NSC 100880): Correlation with clinicalstudies, Cancer Chemother. Rep. 56:515-521 (1972); Gottlieb, J. A. etal., Treatment of malignant melanoma with camptothecin (NSC 100880),Cancer Chemother. Rep. 56:103-105 (1972); and Moertel, C. G., et al.,Phase II study of camptothecin (NSC 100880) in the treatment of advancedgastrointestinal cancer, Cancer Chemother Rep. 56:95-101 (1972).

Despite its potential, interest in CPT as a therapeutic remained at alow ebb until the mid-1980's. By that time, drug therapies were beingevaluated for treating human cancer using human cancer xenograft lines.During these evaluations, human tumors are serially heterotransplantedinto immunodeficient, so-called Anude@ mice, and the mice then testedfor their responsiveness to a specific drug. (Giovanella, B. C., et al.,Cancer 52(7): 1146 (1983)). The data obtained in these studies stronglysupport the validity of heterotransplanted human tumors intoimmunodeficient mammals, such as nude mice, as a predictive model fortesting the effectiveness of anticancer agents.

CPT, and later some of its substituted forms, elicited differentialresponses in the cell cycle of nontumorigenic and tumorigenic humancells in vitro. Although it is not yet understood why CPT and some ofits substituted forms are cytostatic for nontumorigenic cells andcytotoxic for tumorigenic cells, the selective toxicity of the compoundsagainst tumorigenic cells in vitro and in vivo was an especiallyinteresting feature of these drugs.

Investigators began to experiment with various substituted forms of CPT.Good activity was found when various substitutions were made to the CPTscaffold. For example, 9-Amino-20(S)-Camptothecin (9AC) and10,11-Methylendioxy-20(S)-Camptothecin (10,11 MD) are capable of havinghigh anticancer activity against human colon cancer xenografts.Giovanella, B. C., et al., Highly effective topoisomerase-1 targetedchemotherapy of human colon cancer in xenografts, Science 246:1046-1048(1989).

Additionally, 9-nitrocamptothecin (9NC) has shown high activity againsthuman tumor xenograft models. 9NC has a nine position hydrogensubstituted with a nitro moiety. 9NC has inhibited the growth of humantumor xenografts in immunodeficient nude mice and has induced regressionof human tumors established as xenografts in nude mice with little or noappearance of any measurable toxicity. D. Chatterjee et al., Inductionof Apoptosis in Malignant and Camptothecin-resistant Human Cells, 803Annals of the New York Academy of Sciences 143 (1996).

Other substituted CPT compounds that have shown promise include7-ethyl-10-hydroxy CPT, and other 7, 9, 10, 11-substituted compounds.

However, another problem arose when testing began to be done in an invivo environment. CPT compounds contain an α-hydroxy-δ-lactone ringfunctionality that may hydrolyze under physiological conditions. Thelactone moiety may open up easily to yield the carboxylate form,particularly in the presence of human serum albumin (HSA), where 97% of9NC has been observed as converting to the open lactone form. Thomas G.Burke, Chemistry of the Camptothecins in the Bloodstream: DrugStabilization and Optimization of Activity, 803 Annals of the New YorkAcademy of Sciences 29 (1996). As noted above, the biological activityof the closed lactone ring form is far greater than the activity of theopen lactone ring, carboxylated form. In addition, some researchers haveconcluded that a closed lactone ring also may play a role in enhancingpassive diffusion of the CPT molecule into cancer cells. Id.

There have been some attempts to overcome the problems associated withopening of the lactone ring. For example, Published PCT Application WO97/28165 discloses substituted derivatives of camptothecin that areacylated with linear or cyclo alkyl or epoxy moieties at the 20 positionhydroxyl moiety. A stated objective of the acylation is to retain thelactone ring and the 20 position hydroxyl group intact. However, theclass of molecules disclosed suffers from the problem that thepharmacokinetics of release of the active entity, 9-substitutedcamptothecin, are suboptimal. No teaching or suggestion is present inthe WO 97/28165 Application of how to adjust the pharmacokinetics.

Inclusion in liposomes is another solution that has been proposed in thepast to improve the availability of water-insoluble compounds in variousdosage forms. Liposomes are small envelopes of fat-like compoundscontaining an aqueous chamber or chambers within. The water insolublecompound may be dissolved in the fat-like compound comprising theenvelope of the liposome. Liposomes, however, have the disadvantage ofbeing preferentially removed from the circulation and retained in theliver and spleen. This limits their desirability. Liposomes also have anumber of other disadvantages, including potential liposome instabilityand possible change in the size of the liposome upon storage.

There is therefore a need for compositions, methods, apparatus, and kitsthat combine the desirable properties of CPT and its substituted formswith the ability to maintain a closed lactone ring structure.

SUMMARY OF THE INVENTION

The invention relates to compositions comprising a camptothecin and anamorphous cyclodextrin, and to related methods, and apparatus.

DETAILED DESCRIPTION OF THE INVENTION

In an aspect, the invention relates to a composition comprising acamptothecin and an amorphous cyclodextrin. In another aspect, theinvention relates to the composition wherein the camptothecin is asubstituted camptothecin. In still another aspect, the invention relatesto the composition wherein the substituted camptothecin comprises9-nitrocamptothecin, 9-aminocamptothecin,10,11-methylendioxy-20(S)-camptothecin, 7-ethyl-10-hydroxy camptothecin,or another substituted camptothecin that is substituted in at least oneof the 7, 9, 10, 11, or 12 positions. In another aspect, the inventionrelates to the composition of claim 3, wherein the substitutedcamptothecin comprises 9-nitrocamptothecin, or 9-aminocamptothecin.

In a further aspect, the invention relates to the composition whereinsaid amorphous cyclodextrin has a degree of substitution of 2 to 7. Instill another aspect, the invention relates to the composition whereinthe amorphous cyclodextrin is substantially free of pyrogeniccontaminants. In a further aspect, the invention relates to thecomposition wherein the amorphous cyclodextrin comprises hydroxypropyl,hydroxyethyl, glucosyl, maltosyl and maltotriosyl derivatives ofβ-cyclodextrin, carboxyamidomethyl-β-cyclodextrin,carboxymethyl-β-cyclodextrin, sulfobutylether-β-cyclodextrin,hydroxypropyl-β-cyclodextrin or diethylamino-β-cyclodextrin.

In yet another aspect, the invention relates to the composition whereinthe amorphous cyclodextrin comprises hydroxypropyl β-cyclodextrin.

In an aspect, the invention relates to the composition wherein thecyclodextrin comprises hydroxypropyl, hydroxyethyl, glucosyl, maltosyland maltotriosyl derivatives of γ-cyclodextrin. In a further aspect, theinvention relates to the composition wherein the amorphous cyclodextrincomprises a mixture of two or more of α-, β-, or γ-cyclodextrin. Instill another aspect, the invention relates to sterile aqueous solutionscomprising the composition in a form suitable for parenteraladministration.

In an aspect, the invention relates to the composition wherein the ratioof the weight of camptothecin to the weight of cyclodextrin compoundcomprises a range between 1:1 and 1:2000. In a further aspect, theinvention relates to the composition wherein the ratio of the weight ofcamptothecin to the weight of cyclodextrin compound comprises a range ofabout 1:5 to 1:200. In still another aspect, the invention relates tothe composition wherein the ratio of the weight of camptothecin to theweight of cyclodextrin compound comprises a range of about 1:5 to 1:50.

In yet another aspect, the invention relates to the composition whereinthe camptothecin is present in an amount effective to treat undesirableor uncontrolled cell proliferation. In a further aspect, the inventionrelates to the composition wherein the undesirable or uncontrolled cellproliferation comprises restenosis, various cancers, insults to bodytissue due to surgery, diseases that produce fibrosis of tissue,repetitive motion disorders, disorders of tissues that are not highlyvascularized, and proliferative responses associated with organtransplants.

In another aspect, the invention relates to the composition wherein thevarious cancers comprise acute myelogenous leukemia, bladder, breast,cervical, cholangiocarcinoma, chronic myelogenous leukemia, colorectal,gastric sarcoma, glioma, leukemia, lung, lymphoma, melanoma, multiplemyeloma, osteosarcoma, ovarian, pancreatic, prostrate, stomach, ortumors at localized sites including inoperable tumors or in tumors wherelocalized treatment of tumors would be beneficial, and solid tumors. Ina further aspect, the invention relates to the composition wherein thevarious cancers comprise pancreatic or colorectal. In an additionalaspect, the invention relates to the composition wherein the compositionis in a lyophilized form.

In a further aspect, the invention relates to methods of treatingundesirable or uncontrolled cell proliferation comprising administeringthe above-mentioned composition. In another aspect, the inventionrelates to the method wherein the camptothecin is a substitutedcamptothecin. In still another further aspect, the invention relates tothe method wherein the substituted camptothecin comprises9-nitrocamptothecin, 9-aminocamptothecin,10,11-methylendioxy-20(S)-camptothecin, 7ethyl-10-hydroxy camptothecin,or another substituted camptothecin that is substituted at least one ofthe 7, 9, 10, 11, or 12 positions. In yet another aspect, the inventionrelates to the method wherein the substituted camptothecin comprises9-nitrocamptothecin, or 9-aminocamptothecin.

In a further aspect, the invention relates to the method wherein saidamorphous cyclodextrin has a degree of substitution of 2 to 7. In stillanother aspect, the invention relates to the method wherein theamorphous cyclodextrin is substantially free of pyrogenic contaminants.In yet another aspect, the invention relates to the method wherein theundesirable or uncontrolled cell proliferation comprises restenosis,various cancers, insults to body tissue due to surgery, diseases thatproduce fibrosis of tissue, repetitive motion disorders, disorders oftissues that are not highly vascularized, and proliferative responsesassociated with organ transplants. In a further aspect, the inventionrelates to the method wherein the various cancers comprise acutemyelogenous leukemia, bladder, breast, cervical, cholangiocarcinoma,chronic myelogenous leukemia, colorectal, gastric sarcoma, glioma,leukemia, lung, lymphoma, melanoma, multiple myeloma, osteosarcoma,ovarian, pancreatic, prostrate, stomach, or tumors at localized sitesincluding inoperable tumors or in tumors where localized treatment oftumors would be beneficial, and solid tumors. In an aspect, theinvention relates to the method wherein the various cancers comprisepancreatic or colorectal.

In an aspect, the invention relates to implants comprising an implantstructure and the composition. In a further aspect, the inventionrelates to the implant where the implant is a time-release implant. Instill another aspect, the invention relates to the implant where theimplant is a gel or polymer implant. In yet another aspect, theinvention relates to the implant where the implant is coated and thecomposition is contained in the coating. In an aspect, the inventionrelates to the implant where the composition is contained within theimplant structure. In still another aspect, the invention relates to theimplant where the implant is biodegradable or is formed in situ.

In an aspect, the invention relates to a method of treatment comprisinginserting an implant into a body wherein the implant is theabove-mentioned implant. In another aspect, the invention relates to astent comprising the above-mentioned composition.

By forming a complex between camptothecin and cyclodextrins, theinventors believe that camptothecin may be better solubilized in aqueoussolution and the lactone ring of camptothecin may be protected fromhydrolysis. In addition, such complexes of camptothecin:cyclodextrin mayhave reduced ulceration effects when administered to a host. Theinventive compound, compositions comprising the compound, and methods,kits, and apparatus comprising the compound and/or composition will nowbe described in more detail.

An appropriate place to begin describing the invention is to examineputative mechanisms of action because apprehending such mechanisms helpsto put the invention in its proper context. Of course, while such anexplanation is helpful, the inventors do not wish to be bound by aparticular mechanism of action, because complete understanding of suchmechanisms is not necessary to the practice of the invention.

Camptothecin, whether substituted or unsubstituted, is believed tointervene in the mechanism of action of the nuclear enzyme topoisomeraseI (topo I), arresting cells in the S phase. It is believed that CPTaccomplishes this by stabilizing the covalently linked complexes ofDNA-topo I (termed cleavable complexes), thus halting the progression ofreplication forks. This collision of the replication fork with thecleavable complexes is believed to trigger the apoptotic pathway. Z.Darzynkiewicz et al., The Cell Cycle Effects of Camptothecin, 803 Annalsof the New York Academy of Sciences 93 (1996). DNA strand breaks arealso implicated in the cytotoxic effects of CPT. F. Traganos et al.,Induction of Apoptosis by Camptothecin and Topotecan, 803 Annals of theNew York Academy of Sciences 101 (1996).

As discussed above, CPT's activity seems to be diminished for unknownreasons when the lactone ring is opened. HSA seems to be responsiblefor, or at least significantly exacerbates the ring opening. Therefore,stearically protecting the camptothecin molecule by complexing it withan amorphous cyclodextrin so as to prevent the catalytic effects of HSApresumably preserves the in vivo activity of the CPT. Other mechanismsalso may be at work; knowledge of the exact mechanism is not required topractice this invention.

Preferable indications that may be treated using this invention includethose involving undesirable or uncontrolled cell proliferation. Suchindications include restenosis, various types of cancers such as primarytumors, insults to body tissue due to surgery, diseases that producefibrosis of tissue, repetitive motion disorders, disorders of tissuesthat are not highly vascularized, and proliferative responses associatedwith organ transplants.

Specific types of restenotic lesions that can be treated using thepresent invention include coronary, carotid, and cerebral lesions.Specific types of cancers that can be treated using this inventioninclude acute myelogenous leukemia, bladder, breast, cervical,cholangiocarcinoma, chronic myelogenous leukemia, colorectal, gastricsarcoma, glioma, leukemia, lung, lymphoma, melanoma, multiple myeloma,osteosarcoma, ovarian, pancreatic, prostrate, stomach, or tumors atlocalized sites including inoperable tumors or in tumors where localizedtreatment of tumors would be beneficial, and solid tumors. In a morepreferable embodiment, the types of cancer include pancreatic, and/orcolorectal. Treatment of cell proliferation due to insults to bodytissue during surgery may be possible for a variety of surgicalprocedures, including joint surgery, bowel surgery, and cheloidscarring. Diseases that produce fibrotic tissue include emphysema.Repetitive motion disorders that may be treated using the presentinvention include carpal tunnel syndrome. An example of cellproliferative disorders that may be treated using the invention is abone tumor.

The proliferative responses associated with organ transplantation thatmay be treated using this invention include those proliferativeresponses contributing to potential organ rejections or associatedcomplications. Specifically, these proliferative responses may occurduring transplantation of the heart, lung, liver, kidney, and other bodyorgans or organ systems.

The inventive compositions and/or methods may be practiced oradministered by a variety of routes, and may be administered orcoadministered in any conventional dosage form. Coadministration in thecontext of this invention is defined to mean the administration of morethan one therapeutic in the course of a coordinated treatment to achievean improved clinical outcome. Such coadministration may also becoextensive, that is, occurring during overlapping periods of time.

The inventive compounds and/or compositions may be administered orcoadministered orally, parenterally, intraperitoneally, intravenously,intraarterially, transdermally, sublingually, intramuscularly, rectally,transbuccally, intranasally, liposomally, via inhalation, vaginally,intraoccularly, via local delivery (for example by catheter or stent),subcutaneously, intraadiposally, intraarticularly, or intrathecally. Thecompounds and/or compositions according to the invention may also beadministered or coadministered in slow release dosage forms.

By cyclodextrin is meant α-, β-, or γ-cyclodextrin. Cyclodextrins aredescribed in detail in U.S. Pat. No. 4,727,064 to Pitha et al.Cyclodextrins are cyclic oligomers of glucose; these compounds forminclusion complexes with any drug whose molecule can fit into thelipophile-seeking cavities of the cyclodextrin molecule.

If the inventive composition is to be administered parenterally,especially via the intravenous route, the amorphous cyclodextrin ispreferably substantially free of pyrogenic contaminants. Thus thepreferable compositions of matter according to the invention forparenteral administration, especially by the intravenous route, will benonpyrogenic. Nonpyrogenic preparations according to the invention, whenadministered to a subject, preferably do not cause a febrile (basal bodytemperature raising) reaction. Although some bacterial endotoxin may bepresent, the amount is preferably insufficient to elicit a febrilereaction. By substantially pyrogen free is meant that the cyclodextrincontains less than about 10 U.S.P. bacterial endotoxin units per gramusing the method as set forth in United States Pharmacopeia 23 (UnitedStates Pharmacopeial Convention, Rockville, Md., USA). More preferably,the cyclodextrin may contain between 0.1 and 5 U.S.P. bacterialendotoxin units per mg, under conditions specified in the United StatesPharmacopeia 23.

By amorphous cyclodextrin is meant non-crystalline mixtures ofcyclodextrins wherein the mixture is prepared from α-, β-, orγ-cyclodextrin. Preferably, the amorphous cyclodextrin may be preparedby non-selective additions, more preferably by alkylation of the desiredcyclodextrin species. Reactions may be carried out to yield mixturescontaining a plurality of components thereby preventing crystallizationof the cyclodextrin. Various alkylated and hydroxyalkyl-cyclodextrinsmay be made, and of course will vary, depending upon the startingspecies of cyclodextrin and the addition agent used.

Among the amorphous cyclodextrins suitable for compositions according tothe invention are hydroxypropyl, hydroxyethyl, glucosyl, maltosyl andmaltotriosyl derivatives of β-cyclodextrin,carboxyamidomethyl-β-cyclodextrin, carboxymethyl-β-cyclodextrin,sulfobutylether-β-cyclodextrin, hydroxypropyl-β-cyclodextrin anddiethylamino-β-cyclodextrin. In the compositions according to theinvention hydroxy-β-cyclodextrin may be preferable. The substitutedγ-cyclodextrins may also be suitable, including hydroxypropyl,hydroxyethyl, glucosyl, maltosyl and maltotriosyl derivatives ofγ-cyclodextrin.

The cyclodextrin of the compositions according to the invention may beα-, β-, or γ cyclodextrin. α-cyclodextrin contains six glucopyranoseunits; β-cyclodextrin contains seven glucopyranose units; andγ-cyclodextrin contains eight glucopyranose units. The molecule isbelieved to form a truncated cone having a core opening of 4.7-5.3 Å,6.0-6.5 Å and 7.5-8.3 Å in α-, β-, or γ-cyclodextrin respectively. Thecomposition according to the invention may comprise a mixture of two ormore of the α-, β-, or γ-cyclodextrins. More preferably, however, thecomposition according to the invention will comprise only one of the α-,β-, or γ-cyclodextrins.

The particular α-, β-, or γ-cyclodextrin to be used with the particularcamptothecin to form the compositions according to the invention may beselected based on the known size of the camptothecin and the relativesize of the cavity of the cyclodextrin compound. Generally if thecamptothecin is relatively large, a cyclodextrin having a larger cavityis used to make the composition according to the invention. Furthermore,if the camptothecin is administered with an excipient it may bedesirable to use a cyclodextrin compound having a larger cavity in thecomposition according to the invention.

The unmodified α-, β-, or γ-cyclodextrins may be somewhat less desirablein the compositions according to the invention because the unmodifiedforms tend to crystallize and are relatively less soluble in aqueoussolutions. Most preferable for the compositions according to theinvention are the α-, β-, and γ-cyclodextrins that are chemicallymodified or substituted. Chemical substitution at the 2, 3, and 6hydroxyl groups of the glucopyranose units of the cyclodextrin ringsyields increases in solubility of the cyclodextrin compound.

Most preferable as the cyclodextrins in the compositions according tothe invention are amorphous cyclodextrin compounds. By amorphouscyclodextrin is meant non-crystalline mixtures of cyclodextrins whereinthe mixture is prepared from α-, β-, or γ-cyclodextrin. Preferably, theamorphous cyclodextrin is prepared by non-selective alkylation of thedesired cyclodextrin species. Suitable alkylation agents for thispurpose include but are not limited to propylene oxide, glycidol,iodoacetamide, chloroacetate, and 2-diethylaminoethlychloride. Reactionsmay be carried out to yield mixtures containing a plurality ofcomponents thereby potentially preventing crystallization of thecyclodextrin. Various alkylated cyclodextrins may be made, and of coursewill vary, depending upon the starting species of cyclodextrin and thealkylating agent used. Among the amorphous cyclodextrins suitable forcompositions according to the invention are hydroxypropyl, hydroxyethyl,glucosyl, maltosyl and maltotriosyl derivatives of β-cyclodextrin,carboxyamidomethyl-β-cyclodextrin, carboxymethyl-β-cyclodextrin,sulfobutylether-β-cyclodextrin, hydroxypropyl-β-cyclodextrin anddiethylamino-β-cyclodextrin. In the compositions according to theinvention, hydroxypropyl-β-cyclodextrin is preferable although the α- orγ-analogs may also be suitable.

Amorphous hydroxypropyl-β cyclodextrin may be purchased from a number ofvendors including Cerestar, Inc. (Hammond, Ind., USA) under the tradename Encapsin, or from Janssen Pharmaceuticals (Beersen, Belgium). Inaddition, other forms of amorphous cyclodextrin having different degreesof substitution or glucose residue number are available commercially. Amethod for the production of hydroxypropyl-β-cyclodextrin is disclosedin Pitha et. al., U.S. Pat. No. 4,727,064.

The use of cyclodextrins and/or cyclodextrin derivatives have beendisclosed in the following U.S. Pat. Nos.: U.S. Pat. No. 4,024,223 toNoda et al.; U.S. Pat. No. 4,228,160 to Szejtli et al.; U.S. Pat. No.4,232,009 to Hyashi et al.; U.S. Pat. No. 4,351,846 to Matsumoto et al.;U.S. Pat. No. 4,352,793 to Yamahira et al.; U.S. Pat. No. 4,383,992 toLipari; U.S. Pat. No. 4,407,795 to Nicolau; U.S. Pat. No. 4,424,209 toTuttle; U.S. Pat. No. 4,425,336 to Tuttle; U.S. Pat. No. 4,438,106 toWagu et al.; U.S. Pat. No. 4,474,811 to Masuda et al.; U.S. Pat. No.4,478,995 to Shinoda et al.; U.S. Pat. No. 4,479,944 to Hyashi et al.;U.S. Pat. No. 4,479,966 to Hayashi et al.; U.S. Pat. No. 4,497,803 toHarada et al.; U.S. Pat. No. 4,499,085 to Masuda; U.S. Pat. No.4,524,068 to Szejtli et al.; U.S. Pat. No. 4,555,504 to Jones; U.S. Pat.No. 4,565,807 to Uekama et al.; U.S. Pat. No. 4,575,548 to Ueda et al.;U.S. Pat. No. 4,598,070 to Ohwaki et al.; U.S. Pat. No. 4,603,123 toChiesi et al.; U.S. Pat. No. 4,608,366 to Hasegawa et al.; U.S. Pat. No.4,659,696 to Hiari et al.; U.S. Pat. No. 4,623,641 to Szejtili et al.;U.S. Pat. No. 4,663,316 to Ninger et al.; U.S. Pat. No. 4,675,395 toFukazawa et al.; U.S. Pat. No. 4,728,509 to Shimizu et al.; U.S. Pat.No. 4,728,510 to Shibani et al.; and U.S. Pat. No. 4,751,095 to Karl etal.

Camptothecin, when used in the context of this invention, includes bothsubstituted and unsubstituted camptothecins, and analogs thereof. Inparticular, when substituted camptothecins are used, a large range ofsubstitutions may be made to the camptothecin scaffold, while stillretaining activity. In a preferable embodiment, the camptothecinscaffold is substituted at the 7, 9, 10, 11, and/or 12 positions. Suchpreferable substitutions may serve to provide differential activity overthe unsubstituted camptothecin compound. Especially preferable are9-nitrocamptothecin, 9-aminocamptothecin,10,11-methylendioxy-20(S)-camptothecin, 7-ethyl-10-hydroxy camptothecin,or another substituted camptothecin that is substituted in at least oneof the 7, 9, 10, 11, or 12 positions.

Native, unsubstituted, camptothecin can be obtained by purification ofthe natural extract, or may be obtained from the Stehlin Foundation forCancer Research (Houston, Tex.). Substituted camptothecins can beobtained using methods known in the literature, or can be obtained fromcommercial suppliers. For example, 9-nitrocamptothecin may be obtainedfrom SuperGen, Inc. (San Ramon, Calif.), and 9-aminocamptothecin may beobtained from Idec Pharmaceuticals (San Diego, Calif.). Camptothecin andvarious of its analogs may also be obtained from standard fine chemicalsupply houses, such as Sigma Chemicals.

The inventive compositions may include conventional pharmaceuticalexcipients, and other conventional, pharmaceutically inactive, agents.Additionally, the compositions may include more than one camptothecinand more than one cyclodextrin, and/or additional pharmaceuticallyactive agents.

In preferable embodiments, the inventive compositions will contain theactive agents, including the inventive compound, in an amount effectiveto treat an indication of interest. The relative amounts of camptothecincompound and cyclodextrin may vary depending upon the intended use ofthe complex and the effect of the cyclodextrin on the camptothecin.Preferably, the ratio of the weight of camptothecin to the weight ofcyclodextrin compound will be in a range between 1:1 and 1:5000. Withinthis range, the circulating availability of the camptothecin will besignificantly increased when the ratio of the weight of camptothecin tothe weight of cyclodextrin compound is in a range between 1:1 and1:2000. More preferably, the weight to weight ratio of camptothecin tocyclodextrin may be in a range of about 1:5 to 1:200 and even morepreferably in a range of about 1:5 to 1:50.

To produce the formulations according to the invention, a pre-weighedamount of cyclodextrin compound, which is preferably substantiallypyrogen free is placed in a suitable depyrogenated sterile container.Methods for depyrogenation of containers and closure components are wellknown to those skilled in the art and are fully described in the UnitedStates Pharmacopeia 23 (United States Pharmacopeial Convention,Rockville, Md., USA). Generally, depyrogenation is accomplished byexposing the objects to be depyrogenated to temperatures above 4000 Cfor a period of time sufficient to fully incinerate any organic matter.As measured in U.S.P. Bacterial Endotoxin Units, the formulation mightcontain no more than about 10 Bacterial Endotoxin Units per gram ofamorphous cyclodextrin. By substantially pyrogen free is meant that thecyclodextrin contains less than about 10 U.S.P. bacterial endotoxinunits per gram using the U.S.P. method. More preferably, thecyclodextrin will contain between 0.1 and 5 U.S.P. bacterial endotoxinunits per mg, under conditions specified in the United StatesPharmacopeia 23.

Sufficient sterile water for injection may be added to the amorphouscyclodextrin, which is substantially pyrogen free, until the desiredconcentration of cyclodextrin is in solution. To this solution apre-weighed amount of the camptothecin may be added with agitation andwith additional standing if necessary until it dissolves. Excipients, ifany are desired, may be added with or subsequent to adding thecamptothecin.

The solution may then be filtered through a sterile 0.2 micron filterinto a sterile holding vessel and may be subsequently filled in steriledepyrogenated vials and is capped. For products that will be stored forlong periods of time, a pharmaceutically acceptable preservative may beadded to the solution comprising the complex of camptothecin andcyclodextrin prior to filtration, filling and capping, or alternatively,may be added sterilely after filtration.

Additional discussion concerning preparing camptothecin/cyclodextrincompositions according to the invention may be found in V. J. Stella etal., Cyclodextrins: Their Future in Drug Formulation and Delivery, PharmRes, 14(5):556-567 (1997); A. M. Myles et al., Analysis and modelling ofthe structures of beta-cyclodextrin complexes, Biochim Biophys Acta,1199(1):27-36 (1994); H. Arima et al., Enhanced Rectal Absorption andReduced Local Irritation of the Anti-inflammatory Drug Ethyl4-biphenylylacetate in Rats by Complexation with Water-solubleBeta-cyclodextrin Derivatives and Formulation as Oleaginous Suppository,J Pharm Sci, 81(11): 1119-1125 (1992); J. K. Ong et al., Influence ofHydroxypropyl Beta-cyclodextrin on the Stability of Benzylpenicillin inChloroacetate Buffer, J Pharm Pharmacol, 49(6):617-621 (1997); H.Matsuda et al., Inclusion Complexation of P-hydroxybenzoic Acid Esterswith 2-hydroxypropyl-beta-cyclodextrins. On Changes in Solubility andAntimicrobial Activity, Chem Pharm Bull (Tokyo), 41(8):1448-1452 (1993);Y. Nakai et al., Study of the interaction of clobazam with cyclodextrinsin solution and in the solid state, Chem Pharm Bull 38(3):728-732(1990); Y. Watanabe et al., Absorption Enhancement of Polypeptide Drugsby Cyclodextrins. I. Enhanced Rectal Absorption of Insulin FromHollow-type Suppositories Containing Insulin and Cyclodextrins inRabbits, Chem Pharm Bull (Tokyo), 40(11):3042-3047 (1992).

The composition of matter according to the invention may be supplied asa dry powder or as a solution. If the composition of matter is to beinjected into a subject it may be rendered sterile prior to injection.Accordingly, the composition of matter according to the invention may besupplied as a sterile cake, plug or powder or as a sterile Iyophilizedpreparation in a sterile vial suitable for the addition of a sterilediluent, or as a sterile liquid solution in a sterile container.

If the composition is to be administered parenterally, for exampleintravenously, the composition of matter may be rendered sterile priorto such administration. Any of the several known means for renderingsuch pharmaceutical preparations sterile may be used so long as theactive pharmaceutical compound is not inactivated and the complex withthe amorphous cyclodextrin is not degraded. If the active pharmaceuticalcompound is heat stable, the composition of matter according to theinvention may be heat sterilized. If the cytotoxic compound is notheat-stable but is not photo degraded the composition may be sterilizedby exposure to ultraviolet light or by ionizing radiation.Alternatively, the composition of matter if in a powder form may be gassterilized using, for example, ethylene oxide gas. In anotheralternative, the composition of matter according to the invention may befilter-sterilized using a 0.2 micron filter. If the composition ofmatter is an aqueous liquid, it may be filled in a sterile container andsupplied as a sterile liquid ready for further dilution or injectionneat. Alternatively such sterile liquids may be freeze-dried orlyophilized in a sterile container and capped.

Alternatively, the components may be sterilized by any of the knownmethods appropriate to preserving the composition or the camptothecin orthe cyclodextrin prior to mixing in water and may be mixed using sterileequipment and technique. The solution may be lyophilized in sterilecontainers and capped. Prior to use the lyophilized composition ofmatter may be reconstituted using sterile water for injection.

The container closure system used for containing the formulationaccording to the invention may also be treated to remove or destroypyrogenic substances by means known in the art prior to filling andfurther processing. The formulation according to the invention may besupplied as a dry lyophilized powder as mentioned above or as a sterilenon pyrogenic aqueous solution in a sterile container closure systemsuch as a stoppered vial suitable for puncturing with a sterile syringeand needle.

Alternatively the composition according to the invention may be suppliedas a sterile non pyrogenic aqueous solution in a sterile syringe orsyringe and needle. As a sterile solution or powder it may also includea pharmaceutically acceptable preservative. The composition according tothe invention may also be included in other dosage forms in addition tothose appropriate for parenteral administration. Such dosage forms maybe in the form of aqueous suspensions, elixirs, or syrups suitable fororal administration, or compounded as a cream or ointment in apharmaceutically acceptable topical base allowing the inventivecamptothecins to be absorbed across the skin. In addition theformulation according to the invention may be compounded in a lozenge orsuppository suitable for trans mucosal absorption.

Another therapeutically interesting route of administration orcoadministration is local delivery. Local delivery of inhibitory amountsof inventive compounds and/or compositions can be by a variety oftechniques and structures that administer the inventive compounds and/orcompositions at or near a desired site. Examples of local deliverytechniques and structures are not intended to be limiting but rather asillustrative of the techniques and structures available. Examplesinclude local delivery catheters, site specific carriers, implants,direct injection, or direct applications.

Local delivery by a catheter allows the administration of a inventivecompounds and/or compositions directly to the desired site. Examples oflocal delivery using a balloon catheter are described in EP 383 492 A2and U.S. Pat. No. 4,636,195 to Wolinsky. Additional examples of local,catheter-based techniques and structures are disclosed in U.S. Pat. No.5,049,132 to Shaffer et al. and U.S. Pat. No. 5,286,254 to Shapland etal.

Generally, the catheter must be placed such that the inventivecompositions can be delivered at or near the desired site. Dosagesdelivered through the catheter can vary, according to determinationsmade by one of skill, but often are in amounts effective to create acytotoxic or cytostatic effect at the desired site. Preferably, thesetotal amounts are less than the total amounts for systemicadministration of the inventive compositions, and are less than themaximum tolerated dose. The inventive compounds delivered throughcatheters preferably should be formulated to a viscosity that enablesdelivery through a small treatment catheter, and may be formulated withpharmaceutically acceptable additional ingredients (active andinactive).

Local delivery by an implant describes the placement of a matrix thatcontains the inventive compositions into the desired site. The implantmay be deposited by surgery or other means. The implanted matrixreleases the inventive compositions by diffusion, chemical reaction,solvent activators, or other equivalent mechanisms. Examples are setforth in Lange, Science 249:1527-1533 (September, 1990). Often theimplants may be in a form that releases the inventive compositions overtime; these implants are termed time-release implants. The material ofconstruction for the implants will vary according to the nature of theimplant and the specific use to which it will be put. For example,biostable implants may have a rigid or semi-rigid support structure,with inventive composition delivery taking place through a coating or aporous support structure. Other implants may be made of a liquid thatstiffens after being implanted or may be made of a gel. The amounts ofinventive composition present in or on the implant may be in an amounteffective to treat cell proliferation generally, or a specificproliferation indication, such as the indications discussed herein.

One example of local delivery of the inventive composition by an implantis use of a biostable or bioabsorbable plug or patch or similar geometrythat can deliver the inventive composition once placed in or near thedesired site. An example of such implants can be found in U.S. Pat. No.5,429,634 to Narciso, Jr.

A particular application of use of an implant according to the inventionis treatment of cell proliferation in tissue that is not highlyvascularized, as discussed briefly above. An example of such tissue isbone tissue. The difficulty in treating uncontrolled proliferative cellgrowth in bone tissue may be exemplified by the difficulties in treatingbone tumors. Such tumors are typically refractory to treatment, in partbecause bone tissue is not highly vascularized. An implant in or nearthe proliferative site may potentially have localized cytotoxic orcytostatic effects with regard to the proliferative site. Therefore, inone embodiment, the invention may be used to treat bone tumors.

Another example of local delivery by an implant is the use of a stent.Stents are designed to mechanically prevent the collapse and reocclusionof the coronary arteries. Incorporating an inventive composition intothe stent may deliver the inventive composition directly to or near theproliferative site. Certain aspects of local delivery by such techniquesand structures are described in Kohn, Pharmaceutical Technology(October, 1990). Stents may be coated with the inventive composition tobe delivered. Examples of such techniques and structures may be found inU.S. Pat. No. 5,464,650 to Berg et al., U.S. Pat. No. 5,545,208 to Wolffet al., U.S. Pat. No. 5,649,977 to Campbell, U.S. Pat. No. 5,679,400 toTuch, EP 0 716 836 to Tartaglia et al. Alternatively, the inventivecomposition loaded stent may be biorotable, i.e. designed to dissolve,thus releasing the inventive composition in or near the desired site, asdisclosed in U.S. Pat. No. 5,527,337 to Stack et al. The presentinvention can be used with a wide variety of stent configurations,including, but not limited to shape memory alloy stents, expandablestents, and stents formed in situ.

Amounts of the inventive composition delivered by the stent can vary,according to determinations made by one of skill, but preferably are inamounts effective to create a cytotoxic or cytostatic effect at thedesired site. Preferably, these total amounts are less than the totalamounts for systemic administration of the inventive composition, andare preferably less than the maximum tolerated dose. Appropriate releasetimes can vary, but preferably should last from about 1 hour to about 6months, most preferably from about 1 week to about 4 weeks. Formulationsincluding the inventive composition for delivery of the agent via thestent can vary, as determinable by one of skill, according to theparticular situation, and as generally taught herein.

Another example is a delivery system in which a polymer that containsthe inventive composition is injected into the target cells in liquidform. The polymer then cures to form the implant in situ. One variationof this technique and structure is described in WO 90/03768 to Donn.

Another example is the delivery of an inventive composition by polymericendoluminal sealing. This technique and structure uses a catheter toapply a polymeric implant to the interior surface of the lumen. Theinventive composition incorporated into the biodegradable polymerimplant is thereby released at the desired site. One example of thistechnique and structure is described in WO 90/01969 to Schindler.

Another example of local delivery by an implant is by direct injectionof vesicles or microparticulates into the desired site. Thesemicroparticulates may comprise substances such as proteins, lipids,carbohydrates or synthetic polymers. These microparticulates have theinventive composition incorporated throughout the microparticle or overthe microparticle as a coating. Examples of delivery systemsincorporating microparticulates are described in Lange, Science,249:1527-1533 (September, 1990) and Mathiowitz, et al., J. App. PolySci. 26:809 (1981).

Local delivery by site specific carriers describes attaching theinventive composition to a carrier which will direct the drug to thedesired site. Examples of this delivery technique and structure includethe use of carriers such as a protein or non-protein ligand (a steroidreceptor conjugate, for example) or a monoclonal antibody. Certainaspects of these techniques and structures are described in Lange,Science 249:1527-1533.

Local delivery also includes the use of topical applications. An exampleof a local delivery by topical application is applying the inventivecomposition directly to an arterial bypass graft during a surgicalprocedure. Other equivalent examples will no doubt occur to one of skillin the art.

The inventive compositions may be used in the form of kits. Thearrangement and construction of such kits is conventionally known to oneof skill in the art. Such kits may include containers for containing theinventive compositions, and/or other apparatus for administering theinventive compositions.

It will be apparent to those skilled in the art that variousmodifications and variations can be made in the compounds, compositions,kits, and methods of the present invention without departing from thespirit or scope of the invention. Thus, it is intended that the presentinvention cover the modifications and variations of this inventionprovided they come within the scope of the appended claims and theirequivalents. Additionally, the following examples are appended for thepurpose of illustrating the claimed invention, and should not beconstrued so as to limit the scope of the claimed invention.

EXAMPLES Example 1

20(S)-camptothecin is obtained from Sigma. Hydroxypropyl-β-cyclodextrinis obtained according to U.S. Sharma et al., Pharmaceutical and PhysicalProperties of Paclitaxel (Taxol) Complexes with Cyclodextrins, J. Pharm.Sci. 84:1223-30 (1995). A composition containing the 20(S)-camptothecinand the hydroxypropyl-β-cyclodextrin is formed using the proceduresgenerally outlined in U.S. Sharma et al. The complex is then tested forstability according to the methods of U.S. Sharma et al., and foractivity using the human tumor xenograph models outlined in B. C.Giovanella et al., Protocols for the Treatment of Human Tumor Xenograftswith Camptothecins, 803 Annals of the New York Academy of Sciences 181(1996), and Henry S. Friedman et al., Treatment of Central NervousSystem Xenografts with Camptothecins, 803 Annals of the New York Academyof Sciences 210 (1996).

Example 2

9-amino-camptothecin is produced from 20(S)-camptothecin, according toliterature methods. Hydroxypropyl-β-cyclodextrin is obtained accordingto U.S. Sharma et al., Pharmaceutical and Physical Properties ofPaclitaxel (Taxol) Complexes with Cyclodextrins, J. Pharm. Sci.84:1223-30 (1995). A composition containing the 9-amino-camptothecin andthe hydroxypropyl-β-cyclodextrin is formed using the proceduresgenerally outlined in U.S. Sharma et al. The complex is then tested forstability according to the methods of U.S. Sharma et al., and foractivity using the human tumor xenograph models outlined in B. C.Giovanella et al., Protocols for the Treatment of Human Tumor Xenograftswith Camptothecins, 803 Annals of the New York Academy of Sciences 181(1996), and Henry S. Friedman et al., Treatment of Central NervousSystem Xenografts with Camptothecins, 803 Annals of the New York Academyof Sciences 210 (1996).

Example 3

9-aminocamptothecin is produced from 20(S)camptothecin, according toliterature methods. Dimethyl-β-cyclodextrin is obtained according toU.S. Sharma et al., Pharmaceutical and Physical Properties of Paclitaxel(Taxol) Complexes with Cyclodextrins, J. Pharm. Sci. 84:1223-30 (1995).A composition containing the 9-nitrocamptothecin and thedimethyl-β-cyclodextrin is formed using the procedures generallyoutlined in U.S. Sharma et al. The complex is then tested for stabilityaccording to the methods of U.S. Sharma et al., and for activity usingthe human tumor xenograph models outlined in B. C. Giovanella et al.,Protocols for the Treatment of Human Tumor Xenografts withCamptothecins, 803 Annals of the New York Academy of Sciences 181(1996), and Henry S. Friedman et al., Treatment of Central NervousSystem Xenografts with Camptothecins, 803 Annals of the New York Academyof Sciences 210 (1996).

Example 4

9-nitrocamptothecin is produced by nitrating 20(S)camptothecin,according to literature methods. γ-cyclodextrin is obtained according toO. Bekers et al., 2′,3′-Dideoxyinosine (ddI): Its Chemical Stability andCyclodextrin Complexation in Aqueous Media, J Pharm Biomed Anal,11(6):489-493 (June 1993); O. Bekers et al., Effect of Cyclodextrins onthe Chemical Stability of Mitomycins in Alkaline Solution, J PharmBiomed Anal, 9(10-12):1055-1060 (1991); O. Bekers et al., Inclusioncomplexation of doxorubicin and daunorubicin with cyclodextrins, J PharmBiomed Anal, 8(8-12):671-674 (1990). A composition of9-nitrocamptothecin and γ-cyclodextrin is formed using the proceduresgenerally outlined in the O. Bekers et al. articles.

The complex is then tested for stability according to the methodsgenerally outlined in the O. Bekers et al. articles, and for activityusing the human tumor xenograph models outlined in B. C. Giovanella etal., Protocols for the Treatment of Human Tumor Xenografts withCamptothecins, 803 Annals of the New York Academy of Sciences 181(1996), and Henry S. Friedman et al., Treatment of Central NervousSystem Xenografts with Camptothecins, 803 Annals of the New York Academyof Sciences 210 (1996).

Example 5

Irinotecan (Camptothecin-11) is obtained according to Nagahiro Saijo,Clinical Trials of Irinotecan Hydrochloride (CPT, Campto Injection,Topotecin Injection) in Japan, 803 Annals of the New York Academy ofSciences 292 (1996). 3-hydroxypropyl-cyclodextrin is obtained accordingto A. Yoshida et al., Some pharmaceutical properties of3-hydroxypropyl-and 2,3-dihydroxypropyl-beta-cyclodextrins and theirsolubilizing and stabilizing abilities, Chem Pharm Bull (Tokyo),37(4):1059-1063 (1989). A composition of irinotecan and3-hydroxypropyl-cyclodextrin is formed using the procedures generallyoutlined in the A. Yoshida et al. article.

The complex is then tested for stability according to the methodsgenerally outlined in the A. Yoshida et al. article, and for activityusing the human tumor xenograph models outlined in B. C. Giovanella etal., Protocols for the Treatment of Human Tumor Xenografts withCamptothecins, 803 Annals of the New York Academy of Sciences 181(1996), and Henry S. Friedman et al., Treatment of Central NervousSystem Xenografts with Camptothecins, 803 Annals of the New York Academyof Sciences 210 (1996).

Example 6

Topotecan is obtained from Sigma. 2,3-dihydroxypropyl-β-cyclodextrin isobtained according to A. Yoshida et al., Some Pharmaceutical Propertiesof 3-Hydroxypropyl-and 2,3-Dihydroxypropyl-Beta-Cyclodextrins and TheirSolubilizing and Stabilizing Abilities, Chem Pharm Bull (Tokyo),37(4):1059-1063 (1989). A composition of topotecan and2,3-dihydroxypropyl-β-cyclodextrin is formed using the proceduresgenerally outlined in the A. Yoshida et al. article.

The complex is then tested for stability according to the methodsgenerally outlined in the A. Yoshida et al. article, and for activityusing the human tumor xenograph models outlined in B. C. Giovanella etal., Protocols for the Treatment of Human Tumor Xenografts withCamptothecins, 803 Annals of the New York Academy of Sciences 181(1996), and Henry S. Friedman et al., Treatment of Central NervousSystem Xenografts with Camptothecins, 803 Annals of the New York Academyof Sciences 210 (1996).

Example 7

9-nitrocamptothecin is produced by nitrating 20(S)camptothecin,according to literature methods. A series of substitutedbeta-cyclodextrins are prepared, according to F. Hirayama, Developmentand Pharmaceutical Evaluation of Hydrophobic Cyclodextrin Derivatives asModified-Release Drug Carriers, Yakugaku Zasshi, 113(6):425-437 (1993).Compositions including these beta-cyclodextrins and 9-nitrocamptothecinare formed using the procedures generally outlined in the Hirayamaarticle.

These complexes are then tested for stability according to the methodsgenerally outlined in the Hirayama article, and for activity using thehuman tumor xenograph models outlined in B. C. Giovanella et al.,Protocols for the Treatment of Human Tumor Xenografts withCamptothecins, 803 Annals of the New York Academy of Sciences 181(1996), and Henry S. Friedman et al., Treatment of Central NervousSystem Xenografts with Camptothecins, 803 Annals of the New York Academyof Sciences 210 (1996).

Example 8

20(S)-camptothecin is obtained from Sigma. A series of substitutedbeta-cyclodextrins are prepared, according to F. Hirayama, Developmentand Pharmaceutical Evaluation of Hydrophobic Cyclodextrin Derivatives asModified-Release Drug Carriers, Yakugaku Zasshi, 113(6):425-437 (1993).Compositions including these beta-cyclodextrins and 20(S)-camptothecinare formed using the procedures generally outlined in the Hirayamaarticle.

These complexes are then tested for stability according to the methodsgenerally outlined in the Hirayama article, and for activity using thehuman tumor xenograph models outlined in B. C. Giovanella et al.,Protocols for the Treatment of Human Tumor Xenografts withCamptothecins, 803 Annals of the New York Academy of Sciences 181(1996), and Henry S. Friedman et al., Treatment of Central NervousSystem Xenografts with Camptothecins, 803 Annals of the New York Academyof Sciences 210 (1996).

Example 9

The complex obtained in Example 1 is prepared in a gelatin capsule oraldosage formulation. The composition is administered to a patientsuffering from pancreatic adenocarcinoma, according to the protocol setforth in Ethan A. Natelson et al., Phase I Clinical and PharmacologicalStudies of 20-(S)-Camptothecin and 20-(S)-9-Nitrocamptothecin andAnticancer Agents, 803 Annals of the New York Academy of Sciences 224(1996). The amount of the composition administered is normalized toprovide equivalent levels of 9-nitrocamptothecin to those set forth inthe Natelson et al, article. The patient's condition is then monitoredclinically for disease progression.

Example 10

Compositions of 9-nitrocamptothecin and various cyclodextrins areprepared according to Example 8. Differing release rates are determinedfor 9-nitrocamptothecin based upon differing cyclodextrin structuresusing the methods generally set forth in F. Hirayama, Development andPharmaceutical Evaluation of Hydrophobic Cyclodextrin Derivatives asModified-Release Drug Carriers, Yakugaku Zasshi, 113(6):425-437 (1993).Several different compositions having differing release times areobtained. These compositions are then individually compounded intotablets, and are administered orally to human volunteers. Blood samplesare taken from the volunteers, and the amount of closed lactone and openlactone forms of the 9-nitrocamptothecin, together with release time, isdetermined.

Example 11

The composition of Example 1 is prepared. This composition is then usedto prepare a coated stent, using the general teachings of Berg et al.(U.S. Pat. No. 5,464,650). A stenotic lesion is then induced in aconventional animal model, namely a pig artery. The nature anddimensions of the stenotic lesion are then determined using a catheterand an appropriate viewing device. The stent is then deployed at thelesion site, using a conventional stent deployment catheter and balloon.After one week, the pig is sacrificed, and the degree of restenoticgrowth is determined. This amount of growth is compared against acontrol animal where the deployed stent is not coated.

1. A composition comprising a camptothecin and an amorphouscyclodextrin.
 2. The composition of claim 1, wherein the camptothecin isa substituted camptothecin.
 3. The composition of claim 2, wherein thesubstituted camptothecin comprises 9-nitrocamptothecin,9-aminocamptothecin, 10,11-methylendioxy-20(S)-camptothecin,7-ethyl-10-hydroxy camptothecin, or another substituted camptothecinthat is substituted in at least one of the 7, 9, 10, 11, or 12positions.
 4. The composition of claim 3, wherein the substitutedcamptothecin comprises 9-nitrocamptothecin, or 9-aminocamptothecin. 5.The composition of claim 1 wherein said amorphous cyclodextrin has adegree of substitution of 2 to
 7. 6. The composition of claim 1, whereinthe amorphous cyclodextrin is substantially free of pyrogeniccontaminants.
 7. The composition of claim 1, wherein the amorphouscyclodextrin comprises hydroxypropyl, hydroxyethyl, glucosyl, maltosyland maltotriosyl derivatives of β-cyclodextrin,carboxyamidomethyl-β-cyclodextrin, carboxymethyl-β-cyclodextrin,sulfobutylether-β-cyclodextrin, hydroxypropyl-β-cyclodextrin ordiethylamino-β-cyclodextrin.
 8. The composition of claim 7, wherein theamorphous cyclodextrin comprises hydroxypropyl β-cyclodextrin.
 9. Thecomposition of claim 1, wherein the amorphous cyclodextrin compriseshydroxypropyl, hydroxyethyl, glucosyl, maltosyl and maltotriosylderivatives of γ-cyclodextrin.
 10. The composition of claim 1, whereinthe amorphous cyclodextrin comprises a mixture of two or more of α-, β-,or γ-cyclodextrin.
 11. A sterile aqueous solution comprising thecomposition of claim 1 in a form suitable for parenteral administration.12. The composition of claim 1, wherein the ratio of the weight ofcamptothecin to the weight of cyclodextrin compound comprises a rangebetween 1:1 and 1:2000.
 13. The composition of claim 12, wherein theratio of the weight of camptothecin to the weight of cyclodextrincompound comprises a range of about 1:5 to 1:200.
 14. The composition ofclaim 13, wherein the ratio of the weight of camptothecin to the weightof cyclodextrin compound comprises a range of about 1:5 to 1:50.
 15. Thecomposition of claim 1, wherein the camptothecin is present in an amounteffective to treat undesirable or uncontrolled cell proliferation. 16.The composition of claim 15, wherein the undesirable or uncontrolledcell proliferation comprises restenosis, various cancers, insults tobody tissue due to surgery, diseases that produce fibrosis of tissue,repetitive motion disorders, disorders of tissues that are not highlyvascularized, and proliferative responses associated with organtransplants.
 17. The composition of claim 16, wherein the variouscancers comprise acute myelogenous leukemia, bladder, breast, cervical,cholangiocarcinoma, chronic myelogenous leukemia, colorectal, gastricsarcoma, glioma, leukemia, lung, lymphoma, melanoma, multiple myeloma,osteosarcoma, ovarian, pancreatic, prostrate, stomach, or tumors atlocalized sites including inoperable tumors or in tumors where localizedtreatment of tumors would be beneficial, and solid tumors.
 18. Thecomposition of claim 17, wherein the various cancers comprise pancreaticor colorectal.
 19. The composition of claim 1, wherein the compositionis in a lyophilized form.
 20. A method of treating undesirable oruncontrolled cell proliferation comprising administering the compositionof claim
 1. 21. The method of claim 20, wherein the camptothecin is asubstituted camptothecin.
 22. The method of claim 21, wherein thesubstituted camptothecin comprises 9-nitrocamptothecin,9-aminocamptothecin, 10,11-methylendioxy-20(S)-camptothecin,7-ethyl-10-hydroxy camptothecin, or another substituted camptothecinthat is substituted in at least one of the 7, 9, 10, 11, or 12positions.
 23. The method of claim 22, wherein the substitutedcamptothecin comprises 9-nitrocamptothecin, or 9-aminocamptothecin. 24.The method of claim 20, wherein said amorphous cyclodextrin has a degreeof substitution of 2 to
 7. 25. The method of claim 20, wherein theamorphous cyclodextrin is substantially free of pyrogenic contaminants.26. The method of claim 20, wherein the undesirable or uncontrolled cellproliferation comprises restenosis, various cancers, insults to bodytissue due to surgery, diseases that produce fibrosis of tissue,repetitive motion disorders, disorders of tissues that are not highlyvascularized, and proliferative responses associated with organtransplants.
 27. The method of claim 26, wherein the various cancerscomprise acute myelogenous leukemia, bladder, breast, cervical,cholangiocarcinoma, chronic myelogenous leukemia, colorectal, gastricsarcoma, glioma, leukemia, lung, lymphoma, melanoma, multiple myeloma,osteosarcoma, ovarian, pancreatic, prostrate, stomach, or tumors atlocalized sites including inoperable tumors or in tumors where localizedtreatment of tumors would be beneficial, and solid tumors.
 28. Themethod of claim 26, wherein the various cancers comprise pancreatic orcolorectal.
 29. An implant comprising an implant structure and thecomposition of claim
 1. 30. The implant of claim 29, where the implantis a time-release implant.
 31. The implant of claim 29, where theimplant is a gel or polymer implant.
 32. The implant of claim 29, wherethe implant is coated and the composition is contained in the coating.33. The implant of claim 29, where the composition is contained withinthe implant structure.
 34. The implant of claim 29, where the implant isbiodegradable or is formed in situ.
 35. A method of treatment comprisinginserting an implant into a body wherein the implant is the implant ofclaim
 29. 36. A stent comprising the composition of claim
 1. 37. Thestent of claim 36, where the stent is coated and the composition iscontained in the coating.
 38. The stent of claim 36, where thecomposition is contained within the stent structure.
 39. The stent ofclaim 36, wherein the composition is present in an amount effective toreduce undesirable or uncontrolled cell proliferation once the stent isdeployed.
 40. A method of treatment comprising inserting a stent into abody, wherein the stent comprises the composition of claim
 1. 41. Amethod of treatment comprising administering the composition of claim 1through an intraluminal catheter.
 42. A method of treatment comprisingadministering the composition of claim 1 in a local fashion.